摘要
为了探讨细胞角蛋白基因 13(Cytokeratin 13,CK13)在喉癌发生中的作用 ,在CK13基因内部及附近选择 5个微卫星引物进行杂合性丢失 (lossofheterozygosity,LOH)分析 ,于DNA水平间接检测 72例喉鳞状细胞癌患者中该基因的缺失 ,应用NorthernBlot检测 16例喉鳞癌患者的配对肿瘤及癌旁正常组织中CK13基因的表达差异 :同时应用CK13蛋白单克隆抗体对不同分化程度的喉鳞癌组织进行免疫组化染色。结果表明 :5个STR位点均存在LOH ,其中D17S196 4E、D17S2 0 92、D17S791、D17S16 6 5及D17S80 8位点的LOH频率分别为 18.0 3%、2 8.13%、2 7.4 2 %、39.6 8%和 34.85 % ,其中D17S16 6 5位点的LOH频率最高 ,至少一个位点出现LOH的病例高达 77.78% (5 6 / 72 ) ,杂合性丢失与临床分期、淋巴结转移无显著相关 ,但与肿瘤分化程度高低相关 (P <0 .0 5 )。Northernblot结果表明 :16例喉鳞癌患者CK13基因在正常组织中的表达比肿瘤组织显著增强 ,免疫组化结果也显示CK13蛋白在正常组织或高分化肿瘤中的表达明显高于低分化者 ,且存在显著差异 (P <0 .0 1)。证实CK13基因可能在喉鳞状细胞癌的发生中具有重要作用 。
In order to investigate the role of Cytokeratin 13( CK13 ) gene in laryngeal carcinogenesis, we detected the deletion of CK13 gene through LOH analysis indirectly at DNA level using 5 STR primers within and near CK13 gene in 72 cases of laryngeal squamous cell carcinoma, then detected the differential expression between 16 cases of paired normal and cancerous tissue by Northern blot, and performed immunohistochemistry using well characterized monoclonal antibody against CK13 in squamous cell carcinoma of different stages. We found that all of the microsatellite loci exist LOH, and the LOH frequencies were 18.03%? 28.13%? 27.42% ? 39.68% and 34.85% at D17S1964E, D17S2092, D17S791, D17S1665 and D17S808 respectively. The LOH+ cases accounted for 77.78% (56/72), and the frequencies of LOH were not related to the type of laryngeal carcinoma and the lymphoid metastasis; but significantly related to the differentiation, P < 0.05 . CK13 gene is expressed significantly higher in 16 cases of normal tissues than in paired cancerous tissues, and the immunostain revealed that CK13 was expressed in normal laryngeal squamous cell or high differentiation stage, and its expression decreased or disappeared in poor ones, P <0.01. CK13 gene might play an important role in the laryngeal carcinogenesis, acting as a novel tumor suppressor gene, and may be relevant to laryngeal squamous cell carcinoma diagnosis and prognosis. Further research will contribute to conform it.
基金
国家自然科学基金项目 (3 9770 794
3 0 1710 0 8)
辽宁省教育厅基金项目 (2 0 12 10 3 4)
卫生部科学基金项目 (98 1 194) ~~
关键词
喉鳞状细胞癌
细胞角蛋白基因13
杂合性丢失
免疫组化
表达
laryngeal squamous cell carcinoma
Cytokeratin 13 gene
loss of heterozygosity (LOH)
immunohistochemistry
